Part No: MSACL US 2018 Breakfast SeminarIssued year: 2018File size: 3.24mbFile type: pdf
The use of LC/MS analysis in the clinical lab has increased exponentially over the last 10 years. Modern mass spectrometers are extremely sensitive allowing low level detection of many target analytes. However, this sensitivity can come at a price, in that levels of contamination not previously detected with less sensitive instruments can now have larger impact on analysis. The complexity of common matrices such as plasma/serum and urine while presenting different challenges can have a marked influence on method performance.
As a result sample preparation is an extremely important part of the process in order to provide robust methods. This seminar focuses on some of the method development challenges our lab faced when looking at two clinical assays:
Endogenous steroid hormone extraction from serum, and catecholamine extraction from plasma and urine.
Particular emphasis is placed on the various sample preparation options we looked at for the extraction of these analytes. During optimization of the extraction process we investigated analyte recovery, co-extractable matrix components, HPLC column degradation, calibration curve performance and limits of quantitation.
MSACL US 2018 Breakfast Seminar
Part No: PPS451Issued year: 2017File size: 2.99mbFile type: pdf
Whether you need targeted methods for analytes such as Vitamin D metabolites in serum, or methods suitable for extraction of a wide panel of drugs and metabolites from urine, sample preparation before analysis is essential.
This compendium highlights a selection of clinical sample preparation applications from Biotage.
Part No: P177Issued year: 2018File size: 0.58mbFile type: pdf
This poster compares sample preparation options (solid phase extraction using EVOLUTE EXPRESS ABN, and supported liquid extraction using ISOLUTE SLE+) for the extraction of a panel of endogenous steroids from serum.
MSACL NA 2018, Palm Springs, CA
Part No: AN778Issued year: 2013File size: 0.88mbFile type: pdf
This methodology has been designed to give an effective and efficient supported liquid extraction protocol for the clean-up and concentration of salivary cortisol levels. Cortisol is a steroid hormone that when measured from saliva can be used as an indication of stress. Analyte recoveries achieved using this method ranged from 96-99% with RSDs below 3% (n=7) for all analytes.
Cortisol, Saliva, SLE, Supported Liquid Extraction, ISOLUTE, Clinical, Steroid Hormones, Stress
Part No: AN777Issued year: 2013File size: 0.86mbFile type: pdf
Cortisol is a steroid hormone that in urine can be used to diagnose hyper or hypo cortisol diseases such as Cushing’s Syndrome. This methodology has been designed to give an effective and efficient supported liquid extraction protocol for the clean-up and concentration of urinary cortisol levels. Analyte recoveries achieved using this method ranged from 99-101% with RSDs below 5% for all analytes.
Cortisol, Steroid hormone, Cushing's Syndrome, Cushing's Disease, Clinical, SLE, Supported Liquid Extraction, Stress,
Part No: AN891Issued year: 2018File size: 2.35mbFile type: pdf
This application note describes the extraction of a panel of 19 steroid hormones from human serum using EVOLUTE® EXPRESS ABN solid phase extraction plates prior to LC/MS analysis. The simple sample preparation procedure delivers clean extracts and analyte recoveries greater than 80% with RSDs lower than 10% for the majority of analytes. Linearity of >0.99 is achieved for all analytes in the range 5–5000 pg/mL.
Part No: AN890Issued year: 2018File size: 1.73mbFile type: pdf
This application note describes the extraction of a panel of steroid hormones from human serum using ISOLUTE® SLE+ Supported Liquid Extraction plates prior to LC/MS-MS analysis. The simple sample preparation procedure delivers clean extracts and high, reproducible recoveries (>75%, RSD <10% ) for all analytes in human serum, with linearity >0.99 in the range 5–5000 pg/mL.
Part No: AN860Issued year: 2016File size: 0.89mbFile type: pdf
This application note describes a simple supported liquid extraction method for extraction of estrone and estradiol from human serum. An analytical HPLC-MS/MS method that avoids the use of derivatization or fluoride adduct stabilization (two approaches often used to enhance method sensitivity) is used.
High, reproducible analyte recovery and low detection limits (0.001 ng/mL) are achieved.
Part No: AN762Issued year: 2012File size: 0.08mbFile type: pdf
This application note describes the extraction of testosterone and androstenedione from female patient serum samples using ISOLUTE® SLE+ (96 well) plates. This method has been optimized to extract testosterone and androstenedione from female clinical serum samples. The method has achieved very low limits of quantitation for testosterone and androstenedione 0.4nmol/L and 0.9nmol/L respectively. The recoveries for both analytes were greater than 90%.
Testosterone, Androstenedione, Clinical, Serum, ISOLUTE, SLE+, steroids,
Part No: AN740Issued year: 2011File size: 0.38mbFile type: pdf
This application note describes the extraction of testosterone and other steroid hormones from human plasma using ISOLUTE SLE+ supported liquid extraction plates (96-well) with LC-MS/MS analysis. This method describes the use of ISOLUTE SLE+ supported liquid extraction plates (96-well) to extract a range of steroid hormones including testosterone and progesterone from human plasma. This simplified and efficient extraction method has significant analyte recoveries ranging from 90-107% withb LOQs as low as 500 pg/mL.
ISOLUTE, TESTOSTERONE, SLE, SLE+, SUPPORTED LIQUID EXTRACTION, STEROID, HORMONE, PLASMA, ANDROGEN, CLINICAL
Part No: P145Issued year: 2016File size: 0.42mbFile type: pdf
This poster demonstrates a rapid and reliable sample preparation method using Supported Liquid Extraction (ISOLUTE SLE+) to extract a suite of 30 hormone analytes from a hydrolyzed urine matrix. Single injection analysis by LC-MS/MS shows that matrix effects are eliminated by the ISOLUTE SLE+ protocol and that analyte recovery and sensitivity have excellent clinical utility.
Part No: P055Issued year: 2013File size: 0.7mbFile type: pdf
Free urinary cortisol levels can be an indicator of a variety of disorders, such as Cushing Syndrome. This poster presents a simple method for the extraction of cortisol from urine demonstrating good recoveries and low ion suppression. LC-MS/MS analysis was performed using a Waters 2795 Liquid Handling Unit coupled to a Quattro Ultima Pt triple quadrupole MS using electrospray ionization, operated in the MRM mode. Supported liquid extraction method development resulted in recoveries greater than 90% for cortisol spiked urine. Calibration curves constructed using this method from 25-2000 ng/mL, demonstrated excellent linearity with coefficients of determination greater than 0.99.
Cortisol, Stress, Cushin's Syndrome, SLE, Supported Liquid Extraction, SPE, LLEMSACL, San Diego, 2013